Feed additive composition for ruminants

ABSTRACT

Feed additive compositions for ruminants containing (A) at least one member selected from the group consisting of hydrogenated vegetable oil and hydrogenated animal oil each having a melting point of higher than 50° C. and lower than 90° C.; (B) not less than 0.05 wt % and not more than 6 wt % of lecithin; (C) not less than 30 wt % and less than 65 wt % of a biologically active substance; (D) not less than 0.01 wt % and less than 0.8 wt % of a natural vegetable oil; and not less than 0.1 wt % and less than 6 wt % of water exhibit high protection in the rumen and are also superior in dissolution in the lower gastrointestinal tract.

CROSS REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/394,594, filed Apr. 25, 2019, which is a continuation ofInternational Patent Application No. PCT/JP2017/039006, filed on Oct.27, 2017, and claims priority to Japanese Patent Application No.2016-211131, filed on Oct. 27, 2016, both of which are incorporatedherein by reference in their entireties.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to feed additive compositions forruminants. More particularly, the present invention relates to feedadditive compositions for ruminants that are provided with highprotection in the rumen and are superior in dissolution in thegastrointestinal tract.

Discussion of the Background

When ruminants ingest feed, the microorganisms living in the firststomach (rumen) absorb a part of the nutrients in the feed as a nutrientsource. Due to this function, ruminants can absorb, as nutrients,substances that cannot be digested directly. For example, microorganismdigests cellulose to produce saccharides and produces volatile organiccompounds by fermentation with the saccharides. Ruminants absorb suchproducts as nutrition. On the other hand, a nutrient source desired tobe directly absorbed by the ruminant is also digested by themicroorganism and the ruminant can absorb only the substance produced bythe microorganism by fermentation.

To improve the health condition of ruminants and improve theproducibility of the products thereof (e.g., cow's milk, edible meatetc.), it is sometimes desirable to add a nutrient that complementsgeneral feeds.

In such cases, to ensure that the biologically active substance(nutrient) is not ingested by microorganism but effectively absorbed, afeed additive preparation for ruminants that protects the nutrient inthe rumen and makes the nutrient absorbed in the gastrointestinal tractafter the fourth stomach is used.

In feed preparations for ruminants such as lactating cows and the like,a biologically active substance as a nutrient has conventionally beencovered with fat or oil or the like for the purpose of improvingprotection of the biologically active substance in the rumen. Inaddition, a fat or oil having a dissolution promoting effect issometimes used for the purpose of improving dissolution in thegastrointestinal tract and the like. For example, low melting point fatsand oils are easily decomposed by intestinal digestive enzymes such aslipase and the like, and it has been reported that dissolution of abiologically active substance in the gastrointestinal tract can beimproved by adding such fat or oil (see JP-B-49-45224, which isincorporated herein by reference in its entirety). Also, a substanceother than fats and oils is sometimes used. For example, lecithin issometimes used as a promoter of dissolution of a biologically activesubstance from a feed preparation in the gastrointestinal tract ofruminants in view of the action thereof as an emulsifier.

On the other hand, when a ruminant ingests a feed preparation, the feedpreparation stays in the rumen (the first stomach) for a few hours tosome tens of hours, and therefore, a part of the biologically activesubstance is ingested by the microorganism always existing in the rumensuch as protozoa and the like. Dissolution promoters such as lecithinand low melting point fat or oil induce dissolution of biologicallyactive substances in the rumen and, as a result, a problem occurs thatthe protection of the feed preparation in the rumen becomes low.

JP-A-2005-312380, which is incorporated herein by reference in itsentirety, describes a method for producing a dispersion type rumenbypass agent solidified into a sphere with a diameter of 0.5 to 3 mm bya spray granulation method for spraying a mixture containinghydrogenated oil as a protector, lecithin and a monocarboxylic acid saltof an unsaturated or saturated fatty acid having a carbon number of 12to 22 into the air at a melting temperature of the protector (50 to 90°C.). JP-A-2005-312380 discloses that a rumen bypass agent superior inthe protection in the first stomach and releaseability in the fourthstomach is obtained by containing lecithin and stearic acid. On theother hand, JP-A-2005-312380 describes that a rumen bypass agentcontaining 40.0 wt % of L-lysine hydrochloride can be produced by theproduction method. However, the production method described inJP-A-2005-312380 requires use of a mixture having low viscosity forpassage through a spray nozzle, and an L-lysine hydrochloride-containingpreparation having a high content exceeding 40 wt % cannot be obtained.

JP Patent No. 5,040,919, which is incorporated herein by reference inits entirety, describes a dispersion type feed additive composition forruminants, which contains at least one kind of a protector selected froma hydrogenated vegetable oil or hydrogenated animal oil having a meltingpoint of higher than 50° C. and lower than 90° C., lecithin, not lessthan 40 wt % and less than 65 wt % of a basic amino acid and 0.01 to 6wt % of water. This composition contains a high content of not less than40 wt % of a basic amino acid. JP Patent No. 5,040,919 describes thatthe composition has a protection rate improving effect by 0.06 wt % oflecithin and a protection rate improving effect by controlling watercontent, and can achieve a high rumen passage rate.

US-A-2012/244248, which is incorporated herein by reference in itsentirety, describes a ruminant feed additive in which granulated lysinesulfate (particle size 0.3 to 3 mm) is coated with not less than 2layers (desirably not less than 4 layers) of a mixture of hydrogenatedoil and a modifying agent of any of lecithin, stearic acid, oleic acidand palm oil. The content of a lysine sulfate granulation product in thefeed additive is not less than 50% and not more than 60% (not less than37% and not more than 45% based on lysine hydrochloride). The feedadditive contains 0.5 to 10% of a modifying agent and, in the Examplesof US-A-2012/244248, 2 to 4% of a modifying agent is added. When amodifying agent is used, small scratches, cracks and pinholes in thecoating layer of this preparation can be reduced. US-A-2012/244248describes that the rumen bypass rate of the above-mentioned feedadditive is not less than 50% and the small intestine digestion rate isnot less than 70%.

On the other hand, the property of a coating type feed additive markedlydecreases when a ruminant cracks the feed by chewing.

U.S. Pat. No. 8,137,719, which is incorporated herein by reference inits entirety, describes a preparation produced by uniformly mixing afatty acid salt, vegetable oil and lysine hydrochloride and molding theobtained pellets. The preparation contains 15 to 25% of lysine andcontains 1 to 5% of vegetable oil. In the preparation, vegetable oil isused as a liquefiable conditioner, and not only vegetable oil but alsooil, fat, free fatty acid, lipid, lecithin, wax and the like may also beused. U.S. Pat. No. 8,137,719 does not describe a clear role of aliquefiable conditioner, but describes that a fatty acid salt and aliquefiable conditioner form a uniform mixture. The lysine contentdescribed in the Examples of U.S. Pat. No. 8,137,719 is 18.6 to 31%based on lysine hydrochloride and the content cannot be said to be high.

U.S. Pat. No. 8,182,851, which is incorporated herein by reference inits entirety, describes a preparation in which lysine hydrochloride iscoated with a calcium salt of palm oil distillation residue (PFUD) andfatty acid calcium. In the preparation, the calcium salt is dissolvedunder acidic conditions after rumen passage and lysine hydrochloridecontained in the core is eluted. Examples of U.S. Pat. No. 8,182,851describe that a lysine hydrochloride concentration is 20%.

SUMMARY OF THE INVENTION

The aforementioned prior art references disclose that lecithin,vegetable oil, fatty acid and the like are used as liquefiableconditioners, modifying agents or dissolution control agents. While aninfluence of these components on the rumen protection is referred to inthe Examples, an influence on the dissolution in the intestines afterrumen passage is not disclosed.

Accordingly, it is one object of the present invention to provide novelfeed additive compositions for ruminants containing not less than 30 wt% of a biologically active substance, which are of a dispersion typewith less degradation of property due to chewing at feeding time, showhigh protection in the rumen, and are also superior in dissolution inthe gastrointestinal tract.

This and other objects, which will become apparent during the followingdetailed description, have been achieved by the inventors' discoverythat dissolution in the gastrointestinal tract can be enhanced, whilemaintaining high protection in the rumen, by using particular amounts oflecithin and natural vegetable oil.

Therefore, the present invention provides the following.

(1) A feed additive composition for ruminants, comprising

(A) at least one member selected from the group consisting ofhydrogenated vegetable oil and hydrogenated animal oil each having amelting point of higher than 50° C. and lower than 90° C.;

(B) not less than 0.05 wt % and not more than 6 wt % of lecithin;

(C) not less than 30 wt % and less than 65 wt % of a biologically activesubstance;

(D) not less than 0.01 wt % and less than 0.8 wt % of a naturalvegetable oil; and

not less than 0.1 wt % and less than 6 wt % of water.

(2) The composition of (1), wherein the aforementioned (D) is not lessthan 0.1 wt % and not more than 0.4 wt % of a natural vegetable oil.

(3) The composition of (1) or (2), wherein the aforementioned naturalvegetable oil is at least one member selected from the group consistingof soybean oil, palm oil, rape seed oil, canola oil, olive oil, almondoil, avocado oil, and safflower oil.

(4) The composition of any one of (1) to (3), wherein the aforementionednatural vegetable oil comprises an unsaturated fatty acid having acarbon atom number of 18 in not less than 60 wt % and not more than 95wt % relative to the constituent fatty acid of the natural vegetableoil.

(5) The composition of any one of (1) to (4), wherein the aforementionednatural vegetable oil comprises oleic acid in not less than 55 wt % andnot more than 90 wt % relative to the constituent fatty acid of thenatural vegetable oil.

(6) The composition of any one of (1) to (5), wherein the aforementionednatural vegetable oil is olive oil.

(7) The composition of any one of (1) to (6), wherein the aforementionedbiologically active substance is at least one member selected from thegroup consisting of an amino acid, a vitamin and a vitamin-likesubstance.

(8) The composition of any one of (1) to (7), having, as a surfacelayer, a layer substantially free of a biologically active substance.

(9) The composition of (8), wherein the layer substantially free of abiologically active substance has a thickness of not less than 30 μm andnot more than 110 μm.

Effect of the Invention

According to the present invention, a feed additive composition forruminants that is provided with high protection in the rumen and issuperior in dissolution in the gastrointestinal tract can be provided.

The feed additive compositions for ruminants of the present inventioncontaining both lecithin and natural vegetable oil at particularconcentrations can further promote dissolution in the gastrointestinaltract while maintaining high protection in the rumen as compared to whenonly one of lecithin and natural vegetable oil is contained.

According to the feed additive compositions for ruminants of the presentinvention, a large amount of a biologically active substance (e.g.,amino acid etc.) can be efficiently transported up to the smallintestine of lactating cow. Therefore, the lactating cow can absorb alarge amount of the biologically active substance (e.g., amino acidetc.) as a nutrient, as a result of which, for example, it is possibleto increase milk yield production and the like.

BRIEF DESCRIPTION OF THE DRAWINGS

A more complete appreciation of the invention and many of the attendantadvantages thereof will be readily obtained as the same become betterunderstood by reference to the following detailed description whenconsidered in connection with the accompanying drawings, wherein:

FIG. 1 is a graph showing the protection rate, dissolution rate and invitro assumed efficacy rate of the compositions of Examples 1 to 5,Comparative Examples 1 and 2 and control. The left vertical axis showsthe scale of the protection rate and the dissolution rate and the rightvertical axis shows the scale of the in vitro assumed efficacy rate.

FIG. 2 is a graph showing the protection rate, dissolution rate and invitro assumed efficacy rate of the compositions of Examples 6 to 12 andcontrol. The left vertical axis shows the scale of the protection rateand the dissolution rate and the right vertical axis shows the scale ofthe in vitro assumed efficacy rate.

FIG. 3 is a graph plotting in vitro assumed efficacy rates of thecompositions of Examples 6 to 12 and Reference Example relative to theunsaturation rate of a fatty acid having a carbon atom number of 18.

FIG. 4 is an SEM photograph showing a layer of the composition ofExample 1, which is substantially free of a biologically activesubstance.

FIG. 5 is a graph showing the protection rate, dissolution rate and invitro assumed efficacy rate of the compositions of Example 13 andcontrol. The left vertical axis shows the scale of the protection rateand the dissolution rate and the right vertical axis shows the scale ofthe in vitro assumed efficacy rate.

FIG. 6 is a graph showing the protection rate, dissolution rate and invitro assumed efficacy rate of the compositions of Examples 14, 15 andcontrol. The left vertical axis shows the scale of the protection rateand the dissolution rate and the right vertical axis shows the scale ofthe in vitro assumed efficacy rate.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

One of the characteristics of the feed additive composition forruminants of the present invention (to be also referred to as “thecomposition of the present invention” in the following) is that itcontains (A) at least one member selected from the group consisting ofhydrogenated vegetable oil and hydrogenated animal oil each having amelting point of higher than 50° C. and lower than 90° C. (to be alsoreferred to as “Component A” in the following), (B) lecithin (to be alsoreferred to as “Component B” in the following), (C) a biologicallyactive substance (to be also referred to as “Component C” in thefollowing), (D) natural vegetable oil (to be also referred to as“Component D” in the following) and water.

In the present invention, the “feed additive composition for ruminants”generally refers to a composition added to a feed for ruminants andingested when the ruminants ingest the feed. However, it may notnecessarily be added to a feed as long as it is ingested by ruminantsand, for example, the composition of the present invention may beingested by itself by ruminants.

Component A

In the composition of the present invention, component A acts as aprotector. Hydrogenated vegetable oil and hydrogenated animal oil usedas Component A are obtained by solidifying a vegetable oil or animal oilthat is liquid at ordinary temperature (25° C.) by adding hydrogen, andare a concept also including fully hydrogenated oil. The melting pointof the hydrogenated vegetable oil and hydrogenated animal oil used inthe present invention is generally higher than 50° C., and, since theprotection in the rumen may be superior, preferably not less than 55°C., more preferably not less than 60° C. The melting point is generallylower than 90° C., and, since the dissolution in the gastrointestinaltract may be superior, preferably not more than 80° C., more preferablynot more than 70° C. The melting point of the hydrogenated vegetable oiland hydrogenated animal oil used in the present invention is measured bythe open-tube melting point measurement method defined in the JapanAgricultural Standards.

Specific examples of the hydrogenated vegetable oil include soybeanhydrogenated oil, palm hydrogenated oil, rape seed hydrogenated oil,canola hydrogenated oil, olive hydrogenated oil, almond hydrogenatedoil, avocado hydrogenated oil, peanut hydrogenated oil, cottonseedhydrogenated oil, corn hydrogenated oil, safflower hydrogenated oil,sunflower hydrogenated oil, safflower hydrogenated oil, ricehydrogenated oil, candelilla wax, carnauba wax, rice wax, Japan wax,beeswax and the like, preferably soybean hydrogenated oil or soybeanfully hydrogenated oil since they are industrially easily available.Specific examples of the hydrogenated animal oil include beef tallow,lard, whale wax and the like, preferably beef tallow, lard since theyare industrially easily available. These hydrogenated vegetable oil andhydrogenated animal oil may be used singly, or two or more kinds thereofmay be used in combination.

The content of component A in the composition of the present inventiongenerally exceeds 23 wt %, and, since the protection in the rumen may besuperior, is preferably not less than 30 wt %, more preferably not lessthan 35 wt %, relative to the total weight of the composition of thepresent invention. The content is generally less than 60 wt %, and,since a high concentration of a biologically active substance can becontained, preferably not more than 55 wt %, more preferably not morethan 50 wt %, relative to the total weight of the composition of thepresent invention.

Component B

Lecithin used as Component B is considered to act as an emulsifier, tomodify the surface of the biologically active substance and uniformlydisperse the active substance in the molten protector without unevenlydistributing the substance.

Specific examples of lecithin include plant-derived lecithins such assoybean lecithin, rape lecithin, rapeseed lecithin, sunflower lecithin,safflower lecithin, cottonseed lecithin, corn lecithin, linseedlecithin, sesame lecithin, rice lecithin, coconut lecithin, palmlecithin and the like; egg-yolk lecithin and the like, preferablyplant-derived lecithin, more preferably soybean lecithin, since they areindustrially easily available. These lecithins may be, for example,hydrogenated product, enzyme treatment product, enzyme decompositionproduct, lecithin fractionated product or the like. These lecithins maybe used singly, or two or more kinds thereof may be used in combination.

The content of Component B in the composition of the present inventionis generally not less than 0.05 wt % and, since the protection in therumen could be superior, preferably not less than 0.5 wt %, morepreferably not less than 1 wt %, relative to the total weight of thecomposition of the present invention. The content is generally not morethan 6 wt % and, since the protection in the rumen could be superior,preferably not more than 5 wt %, more preferably not more than 3 wt %,particularly preferably not more than 2 wt %, relative to the totalweight of the composition of the present invention.

Component C

The biologically active substance used as Component C is notparticularly limited as long as it is a substance capable of exhibitinga bioactive function in vivo when ingested by ruminants. For example, anamino acid, vitamin, vitamin-like substance, enzyme, protein, peptideand the like can be mentioned. From the aspect of probiotics, it ispreferably an amino acid, vitamin or a vitamin-like substance.

The amino acid may be a free amino acid, or a physiologically acceptablesalt. Examples of the physiologically acceptable salts of the amino acidinclude salts with inorganic bases, salts with inorganic acids and saltswith organic acids and the like. Examples of the salt with an inorganicbase include salts with alkali metals such as sodium, potassium, lithiumand the like, salts with alkaline earth metals such as calcium,magnesium and the like, ammonium salt and the like. Examples of the saltwith an inorganic acid include salts with hydrohalic acid (hydrochloricacid, hydrobromic acid, hydroiodic acid etc.), sulfuric acid, nitricacid, phosphoric acid and the like. Examples of the salt with an organicacid include salts with formic acid, acetic acid, propionic acid, oxalicacid, succinic acid, maleic acid, fumaric acid, citric acid and thelike. Any of L-form, D-form and DL-form of the amino acid can be used,and preferred is the L-form or DL-form, and further preferred is theL-form.

Specific examples of amino acid include basic amino acids such asarginine, histidine, lysine, hydroxylysine, ornithine, citrulline andthe like or a physiologically acceptable salt thereof; neutral aminoacids such as glycine, alanine, valine, leucine, isoleucine, serine,threonine, asparagine, glutamine, tryptophan, 5-hydroxytryptophan,cystine, cysteine, methionine, proline, hydroxyproline, phenylalanine,tyrosine and the like or a physiologically acceptable salt thereof; andacidic amino acids such as aspartic acid, glutamic acid and the like ora physiologically acceptable salt thereof and the like. The amino acidis preferably a basic amino acid or a physiologically acceptable saltthereof since it shows high physiologically activating function on dairycattle. Among others, the amino acid is more preferably lysine or aphysiologically acceptable salt thereof, particularly preferably a saltof lysine with an inorganic acid, most preferably lysine hydrochloride,since it is considered to be most important for increasing the milkyield of lactating cow. These amino acids may be used alone, or two ormore kinds thereof may be used in combination.

The amino acid and a physiologically acceptable salt thereof may be anyof those obtained by extraction and purification from naturallyoccurring animals, plants and the like, or those obtained by chemicalsynthesis method, fermentation method, enzymatic method or generecombination method may be used. Alternatively, a commerciallyavailable product may be used as it is or after pulverizing. When theamino acid is pulverized, the particle size thereof is preferably notmore than 100 μm, more preferably not more than 75 μm.

Specific examples of vitamin include water-soluble vitamins such asvitamin B₁, vitamin B₂, vitamin B₆, vitamin B₁₂, folic acid, niacin,pantothenic acid, biotin, vitamin C and the like. These vitamins may beused alone, or two or more kinds thereof may be used in combination, andan appropriate one may be selected.

The vitamin-like substance refers to a compound having a physiologicalaction similar to that of vitamin and is biosynthesizable in the bodyunlike vitamin. Concrete examples thereof include choline (e.g., cholinechloride, CDP (cytidine diphosphate)choline, choline bitartrate etc.),p-aminobenzoic acid, lipoic acid, carnitine, orotic acid, ubiquinone andthe like. These vitamin-like substances may be used alone, or two ormore kinds thereof may be used in combination.

The content of component C in the composition of the present inventionis generally not less than 30 wt % and, since a large amount of abiologically active substance can be efficiently afforded, preferablynot less than 35 wt %, more preferably not less than 40 wt %, relativeto the total weight of the composition of the present invention. Thecontent is generally less than 65 wt % and, since protection in therumen could be superior, preferably not more than 60 wt %, relative tothe total weight of the composition of the present invention.

Component D

The natural vegetable oil used as component D refers to a vegetable oilwhich is liquid at ordinary temperature (25° C.), and is a conceptdistinguished from the hydrogenated vegetable oil possibly used forcomponent A.

Specific examples of the natural vegetable oil include soybean oil, palmoil, rape seed oil, canola oil, olive oil, almond oil, avocado oil,safflower oil, sunflower oil, corn oil, rice oil and the like. Preferredare soybean oil, palm oil, rape seed oil, canola oil, olive oil, almondoil, avocado oil and safflower oil. These natural vegetable oils may beused alone, or two or more kinds thereof may be used in combination.These natural vegetable oils may be subjected to a treatment such astransesterification, partition treatment and the like as long as it isliquid at ordinary temperature.

While the kind of the fatty acid (constituent fatty acid) constitutingComponent D is not particularly limited, for example, saturated orunsaturated fatty acids having a carbon atom number of 12 to 24 such aslauric acid, myristic acid, palmitic acid, pulmitoleic acid, stearicacid, oleic acid, linoleic acid, linolenic acid, arachidic acid, behenicacid and the like can be mentioned. From the aspect of dissolution inthe gastrointestinal tract, component D preferably includes unsaturatedfatty acids having a carbon atom number of 18 such as oleic acid,linoleic acid, linolenic acid and the like.

While the constitution rate of saturated and unsaturated fatty acids inComponent D is not particularly limited, component D preferably containsnot less than 60 wt % and not more than 95 wt %, more preferably notless than 80 wt % and not more than 95 wt %, of unsaturated fatty acidhaving a carbon atom number of 18, relative to the constituent fattyacid of component D, since the in vitro assumed efficacy rate could besuperior.

Component D preferably contains not less than 55 wt % and not more than90 wt %, more preferably not less than 70 wt % and not more than 90 wt%, of oleic acid relative to the constituent fatty acid of Component D.Specific examples of component D containing not less than 55 wt % andnot more than 90 wt % of oleic acid relative to the constituent fattyacid include olive oil and the like.

While the unsaturation rate of fatty acid having a carbon atom number of18 contained in Component D (rate of weight of unsaturated fatty acidhaving a carbon atom number of 18 to the total weight of saturated fattyacid and unsaturated fatty acid having a carbon atom number of 18) isnot particularly limited, it is generally not less than 40% and, sincethe in vitro assumed efficacy rate could be superior, preferably notless than 50%, more preferably not less than 55%. While the upper limitof the unsaturation rate is not particularly limited, it is generally100%.

The content of Component D in the composition of the present inventionis generally not less than 0.01 wt % and, since the in vitro assumedefficacy rate could be superior, preferably not less than 0.05 wt %,more preferably not less than 0.1 wt %, relative to the total weight ofthe composition of the present invention. The content is generally lessthan 0.8 wt % and, since the in vitro assumed efficacy rate could besuperior, preferably not more than 0.6 wt %, more preferably not morethan 0.4 wt %, particularly preferably less than 0.4 wt %, relative tothe total weight of the composition of the present invention.

Water contained in the composition of the present invention isconsidered to affect the preservation stability of the composition ofthe present invention and improve protection in the rumen. Watercontained in the composition of the present invention is notparticularly limited as long as it is generally used for producing feedadditive compositions and, for example, ultrapure water, pure water, ionexchange water, distilled water, purified water, tap water and the likecan be mentioned.

The content of water (water content) in the composition of the presentinvention is generally not less than 0.1 wt % and, since the protectionin the rumen could be superior, preferably not less than 2 wt %,relative to the total weight of the composition of the presentinvention. The content is generally less than 6 wt % and, since theprotection in the rumen could be superior, preferably not more than 5 wt%, more preferably not more than 4 wt %, relative to the total weight ofthe composition of the present invention.

The water content of the composition of the present invention can bedetermined by measuring an amount of decrease after heating at 105° C.for 20 min by Kett moisture analyzer (infrared Moisture Balance FD-610).

The composition of the present invention may contain, besides ComponentsA to D and water, other component other than those. Such othercomponents are not particularly limited as long as the object of thepresent invention is not impaired. For example, excipients such ascalcium carbonate, silicon dioxide and the like; lubricants such asmagnesium stearate, calcium stearate, talc and the like; pH adjusterssuch as sodium hydrogen carbonate, citric acid and the like; anticakingagents such as calcium silicate, sodium aluminosilicate and the like;and the like can be mentioned. Such other component may be used singly,or two or more kinds thereof may be used in combination.

The composition of the present invention is preferably formed in a shapeeasily ingestable for ruminants. While the shape is not particularlylimited, for example, spherical, granular, pellet shape, rugby ballshape, pressed barley shape, hen's egg shape and the like can bementioned.

It is preferable that the composition of the present invention has aspherical or a shape similar thereto. While the particle size of amolded product of the composition of the present invention is notparticularly limited, it is generally 0.1 to 20 mm and, from the aspectof mixing level with a feed, preferably 0.3 to 10 mm, more preferably0.5 to 5 mm. The particle size of the composition of the presentinvention is defined by sieve analysis using the standard sieve definedin JIS Z 8801 of Japanese Industrial Standards, which is incorporatedherein by reference in its entirety.

The production method of the composition of the present invention is notparticularly limited, and the composition of the present invention maybe produced by a method known per se. For example, it can be produced bythe method described in WO2008/041371, US-A-2009/0232933, WO2009/122750,US-A-2011/0081444, which are incorporated herein by reference in theirentireties, or a method analogous thereto. Specifically, the compositionof the present invention can be produced by a method includingsolidifying a molten mixture containing Components A to D by immersingin water or the like.

A preparation method of the molten mixture containing Components A to Dis not particularly limited and, for example, a method including heatingComponents A to D (optionally containing other component when desired)using a commercially available extruder (preferably, twin screwextruder) and the like, and the like can be mentioned. The order ofaddition of Components A to D to a cylinder of the extruder is notparticularly limited. To coat the surface of Component C with ComponentB, the Components B and C may be mixed with a Nauta mixer or the likebefore addition, or Components A to D may be added almost simultaneouslyto increase production efficiency. Alternatively, the molten mixture canalso be obtained by mixing Components A and C in advance at around roomtemperature, adding the remaining components and heating the mixture.Component C may be pulverized before use. For example, Component C maybe used after pulverizing with a pulverizer to a particle size ofpreferably not more than 100 μm, more preferably not more than 75 μm,and sieving as necessary.

The temperature at which Components A to D are heated is notparticularly limited as long as it is not less than the melting point ofComponent A. It is preferably 5 to 15° C. higher than the melting pointof Component A. For example, when soybean fully hydrogenated oil(melting point: 67 to 71° C.) is used as Component A, it is heated at 80to 85° C. In this case, the component other than Component A is notnecessarily melted. For example, when L-lysine hydrochloride (meltingpoint: 263° C.) is used as component C, L-lysine hydrochloride may bedispersed without melting and the molten mixture may be in a slurrystate. It is not necessary to heat at a temperature not less than themelting point of Component A from the beginning of heating. A stablemolten mixture is obtained efficiently by, for example, first preheatingmaterials at a temperature 5 to 10° C. lower than the melting point ofComponent A, then conveying the materials by a screw in the cylinder ofthe extruder, and then heating them at a predetermined temperature notless than the melting point of Component A.

The instrument that can be utilized for preparing the molten mixture isnot limited to the extruder, and any instrument may be usedappropriately as long as it can prepare a molten mixture that can becomea droplet when dropped naturally.

A method for immersing the molten mixture containing Components A to Din water is not particularly limited. For example, a method includingretaining the molten mixture in a container having a hole (pore) with apredetermined diameter and dropping the molten mixture into water fromthe hole, and the like can be mentioned. When the molten mixture isdropped (preferably, free fall) from the hole with a predetermineddiameter, it is cleaved by the action of surface tension during droppingto become respectively independent droplets. When the droplet is droppedinto a water tank at a given temperature, the droplet is instantaneouslycooled in water to solidification and a solid with a given shape isobtained. When the droplet solidifies into solid, the water in the watertank is taken into the solid. This water can be decreased by aheat-drying treatment (described later). When the molten mixture isimmersed in water, a part of the biologically active substance may bedissolved in water; however, the amount thereof is extremely small.

The diameter of the hole in the container retaining the molten mixturemay be appropriately selected according to the size of thefinally-obtained solid (solidified droplet of the molten mixture). Forexample, when a solid with a particle size of about 3 to 5 mm isproduced, the diameter of the hole only needs to be set to 0.5 to 3 mm,and when a solid with a particle size of about 5 to 10 mm is produced,the diameter of the hole only needs to be set to 3 to 5 mm. The diameterof the hole in the container for storing a molten mixture is generally0.5 to 5 mm, preferably 1 to 4 mm.

While the container retaining the molten mixture is not particularlylimited as long as it has a hole with a predetermined diameter, amulti-hole shooter is preferably used since it can efficiently increasethe production amount. Here, the “multi-hole shooter” refers to acontainer having a plurality of perforations in the bottom and afacility for temporarily retaining the molten mixture. The container forretaining the molten mixture is preferably provided with a heat facilityto prevent cooling of the molten mixture to be retained.

The drop distance (e.g., distance from the bottom surface of themulti-hole shooter to water surface) of the molten mixture is notparticularly limited and it is generally 10 mm to 1.5 m, preferably 30mm to 1.0 m. The shape of the finally-obtained solid can be changed byadjusting the drop distance of the molten mixture. For example, when amolten mixture heated to about 65° C. is dropped into water, a dropdistance of 50 to 150 mm affords a solid with a spherical shape to arugby ball-like shape. A longer drop distance leads to a large impactenergy with the water surface, and a solid with a shape of flattenedpressed barley is obtained. For example, when the drop distance is about0.5 m, a solid with a shape of pressed barley with an undulating fringeis obtained.

The temperature of the molten mixture when dropped into water is notparticularly limited and is generally 60 to 90° C., and preferably 70 to90° C. in view of the melting point of component A and the like.

The temperature of the water into which the molten mixture is dropped isnot particularly limited as long as the molten mixture isinstantaneously solidified, and is generally 0 to 30° C. When thetemperature of water into which the molten mixture is dropped is toohigh, the particle shape of the obtained solid tends to collapse, turninto a flake and break easily. The water temperature is preferablymaintained constant. For example, the temperature of water into whichthe molten mixture is dropped can be maintained constant by continuoussupplementation of water at a predetermined temperature and the like.

A method for collecting a mixture solidified in water is notparticularly limited. When the water temperature is to be maintainedconstant by continuously supplementing water, the solidified mixture(specific gravity: about 1.1) may be collected using a net, a netcontainer or the like.

When the composition of the present invention is produced by a methodincluding solidifying a molten mixture containing Components A to D byimmersing in water, the method preferably further includes applying aheat-drying treatment to the solidified mixture. The water content ofthe composition of the present invention can be controlled by theheat-drying treatment. The heat-drying treatment can be performed, forexample, by exposing the solidified mixture generally for severalminutes to several tens of minutes to an atmosphere (e.g., hot water,vapor, hot air etc.) set to a temperature lower than the melting pointof Component A contained in the solidified mixture, or the like. Thetime of the heat-drying treatment can be appropriately determined basedon the temperature of the heat-drying treatment, the kind of ComponentA, the amount of the solidified mixture and the like. For example, thesolidified mixture may be exposed for a long time (e.g., 0.5 to 2 hretc.) to an atmosphere set to a temperature lower than the melting pointof Component A contained in the solidified mixture.

The composition of the present invention preferably has a layersubstantially free of a biologically active substance as the surfacelayer thereof. The composition of the present invention may havewater-repellency by the presence of a layer substantially free of abiologically active substance as the surface layer. As used herein, the“layer substantially free of a biologically active substance” meanseither a layer completely free of a biologically active substance or alayer containing a biologically active substance in an amount notinhibiting water-repellency (generally not more than 2 wt %, preferablynot more than 1 wt %).

The thickness of the layer substantially free of a biologically activesubstance is generally not less than 30 μm and not more than 110 μm and,since water-repellency could be superior, preferably not less than 30 μmand not more than 80 μm.

The composition of the present invention having a layer substantiallyfree of a biologically active substance as the surface layer can beproduced by, for example, the aforementioned production method, that is,a method including solidifying a molten mixture containing Components Ato D by immersing in water and subjecting the solidified mixture to aheat-drying treatment or the like. The layer substantially free of abiologically active substance is considered to be formed because thebiologically active substance on the surface is dissolved in water whenthe molten mixture is immersed in water and then the surface of thesolidified mixture is smoothened by the heat treatment.

The protection in the rumen and dissolution in the gastrointestinaltract of the composition of the present invention can be evaluated bythe following method.

In the following method, the concentration of the biologically activesubstance in the test solution is measured by liquid chromatography(manufactured by Hitachi). When the biologically active substance islysine, it can also be measured by a biosensor (manufactured by OjiScientific Instruments).

Measurement of Concentration (Concentration A) of Biologically ActiveSubstance for Calculation of Protection Rate

Using a dissolution tester (manufactured by TOYAMA SANGYO CO., LTD.), apreparation sample (about 3 g) is placed in ultrapure water (900 ml)(produced using Milli Q (manufactured by Millipore)) heated to atemperature (e.g., 39° C.) corresponding to the body temperature ofruminants (e.g., dairy cattle etc.), and the mixture is stirred at 100rpm. At 20 hr from the start of stirring, 2 ml of the stirring testsolution is collected for protection rate measurement, and theconcentration of the biologically active substance is measured(concentration A, unit: mg/dl).

Measurement of Concentration (Concentration B) of Biologically ActiveSubstance for Calculation of Dissolution Rate

To the test solution immediately after collection of the above-mentionedsample for protection rate measurement is added with stirring at 100 rpman aqueous solution (8 ml) of a bile powder (manufactured by Wako PureChemical Industries, Ltd.) and pancreatin (manufactured by Wako PureChemical Industries, Ltd.) (concentration of bile powder and pancreatinis 23.4 g/100 ml for each) to give a small intestine-corresponding testsolution. At 5 hr from the addition of the aqueous solution, 2 ml of thestirring test solution is collected for dissolution rate measurement,and the concentration of the biologically active substance is measured(concentration B, unit: mg/dl).

Calculation of Protection Rate and Dissolution Rate of BiologicallyActive Substance

The protection rate and dissolution rate of biologically activesubstance are calculated by the following formulas.

protection rate[0]={1−(concentration A[mg/dl]×9.08)/(preparation sampleweight[g]×1000×content of biologically active substance in preparationsample [wt %]/100)}×100

dissolution rate[%]={((concentration B[mg/dl]−concentrationA[mg/dl])×9.02)/(preparation sample weight[g]×1000×content ofbiologically active substance in preparation sample [wt %]/100)}×100

The in vitro assumed efficacy rate of the composition of the presentinvention can be calculated from the following formula.

in vitro assumed efficacy rate[%]=(dissolution rate [%])×(content[wt %]of biologically active substance)/100

The ruminants for which the composition of the present invention is usedare not particularly limited. For example, bovine, sheep, goat, deer,giraffe, camel and llama and the like can be mentioned. Preferred isbovine.

The amount of the composition of the present invention to be added to afeed for ruminants is not particularly limited, and can be appropriatelyadjusted according to the necessary amount of the biologically activesubstance and the like. The composition of the present invention isgenerally added to a feed and used together with the feed to be ingestedby the ruminants. However, as long as the composition is ingested by theruminants, it may not necessarily be added to a feed. For example, thecomposition of the present invention can be ingested by itself theruminants.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES Experimental Example 1. Study of Content of Natural VegetableOil Example 1

Soybean fully hydrogenated oil (manufactured by Yokozeki Oil & FatIndustries Co., Ltd., melting point: 67° C.), soybean lecithin(manufactured by ADM, Yelkin TS), L-lysine hydrochloride (manufacturedby Ajinomoto Co., Inc.) and olive oil (manufactured by JOM, extravirgin) in the proportions shown in the following Table 1 werecontinuously cast into a twin screw extruder (manufactured by CosmotecCo., Ltd.).

Thereafter, the mixture was heated (preheating temperature: 65° C., mainheating temperature: 85° C., set temperature for outlet: 70° C.), meltedand mixed in a cylinder to give a molten mixture in a molten slurrystate. The obtained molten mixture was discharged from the outlet of theextruder, cast into a multi-hole shooter (number of holes: 2060, holediameter: 2 mm), and the molten mixture was freely dropped from the holeof a multi-hole shooter into the water tank for cooling (watertemperature: 5 to 15° C.). The distance from the multi-hole shooter tothe water surface of the water tank for cooling was 10 cm. The moltenmixture that dropped from the multi-hole shooter became droplet duringdropping, immersed in water, cooled and solidified instantaneously. Theattached water was dehydrated by blowing at room temperature, and thesolid was subjected to a heat-drying treatment by a fluidized-bed dryer(manufactured by Ajinomoto Co., Inc.) set to 52° C. for 7 min to givegranules (feed additive composition for ruminants). In the following,the granulated product is referred to as the composition of Example 1.

Examples 2-5, Comparative Examples 1 and 2, and Control

In the same manner as in Example 1 except that soybean fullyhydrogenated oil (manufactured by Yokozeki Oil & Fat Industries Co.,Ltd., melting point:67° C.), soybean lecithin (manufactured by ADM,Yelkin TS), L-lysine hydrochloride (manufactured by Ajinomoto Co., Inc.)and olive oil (manufactured by JOM, extra virgin) were cast in theproportions shown in the following Table 1, feed additive compositionsfor ruminants of Examples 2 to 5, Comparative Examples 1 and 2, and thecontrol (hereinafter to be referred to as composition of Examples 2 to5, Comparative Examples 1 and 2, and the control).

The lysine content, water content, protection rate and dissolution rateshown in Examples 1 to 5, Comparative Examples 1 and 2, and the controlwere measured in the following manner.

Measurement of Lysine Content of Feed Additive Composition

The lysine content of each composition was measured using a biosensor(manufactured by Oji Scientific Instruments).

Measurement of Water Content of Feed Additive Composition

The water content of each composition was determined by measuring, byKett moisture analyzer (infrared Moisture Balance FD-610), an amount ofdecrease after heating at 105° C. for 20 min.

Measurement of Protection Rate and Dissolution Rate

The concentration of the biologically active substance in the followingtest solutions was measured using a biosensor (manufactured by OjiScientific Instruments).

Measurement of Concentration (Concentration A) of Biologically ActiveSubstance (L-Lysine) for Calculation of Protection Rate

Using a dissolution tester (manufactured by TOYAMA SANGYO CO., LTD.), apreparation sample (about 3 g) was placed in ultrapure water (producedusing Milli Q (manufactured by Millipore)) (900 ml) heated to 39° C.corresponding to the body temperature of dairy cattle, and the mixturewas stirred at 100 rpm. At 20 hr from the start of stirring, 2 ml of thestirring test solution was collected for protection rate measurement,and the concentration of the biologically active substance (L-lysine)was measured (concentration A, unit: mg/dl).

Measurement of Concentration (Concentration B) of Biologically ActiveSubstance (L-Lysine) for Calculation of Dissolution Rate

To the test solution immediately after collection of the above-mentionedsample for protection rate measurement was added with stirring at 100rpm an aqueous solution (8 ml) of a bile powder (manufactured by WakoPure Chemical Industries, Ltd.) and pancreatin (manufactured by WakoPure Chemical Industries, Ltd.) (concentration of bile powder andpancreatin is 23.4 g/100 ml for each) to give a smallintestine-corresponding test solution. At 5 hr from the addition of theaqueous solution, 2 ml of the stirring test solution was collected fordissolution rate measurement, and the concentration of the biologicallyactive substance (L-lysine) was measured (concentration B, unit: mg/dl).

Calculation of Protection Rate and Dissolution Rate of BiologicallyActive Substance (L-Lysine)

The protection rate and dissolution rate of biologically activesubstance (L-lysine) were calculated by the following formulas.

protection rate(%)={1−(concentration A[mg/dl]×9.08)/(preparation sampleweight[g]×1000×content of biologically active substance in preparationsample [wt %]/100)}×100

dissolution rate(%)={((concentration B[mg/dl]−concentration A[mg/dl])×9.02)/(preparation sample weight[g]×1000×content ofbiologically active substance in preparation sample [wt %]/100)}×100

The in vitro assumed efficacy rates of the compositions of Examples 1 to5, Comparative Examples 1 and 2, and the control were calculated fromthe following formula.

in vitro assumed efficacy rate[%]=(dissolution rate [%])×(content[wt %]of biologically active substance)/100

The L-lysine content, water content, protection rate, dissolution rateand in vitro assumed efficacy rate of the compositions of Examples 1 to5, Comparative Examples 1 and 2, and the control are shown in thefollowing Table 1. In addition, the protection rate, dissolution rateand in vitro assumed efficacy rate of the compositions of Examples 1 to5, Comparative Examples 1 and 2, and the control are shown in FIG. 1.

The contents of soybean fully hydrogenated oil, soybean lecithin andolive oil in the molten mixture do not change before and aftergranulation in water.

TABLE 1 blending proportion [wt %] L-lysine water (A) content contentsoybean (C) [wt %] of [wt %] of in vitro fully (B) L- (D) feed feedprotection dissolution assumed hydrogenated soybean lysine oliveadditive additive rate rate efficacy oil lecithin hydrochloride oilcomposition composition [%] [%] rate [%] control 41.9 1.1 57 — 37.0 4.5100 37.7 13.9 Ex. 1 41.7 1.1 57 0.2 42.5 2.1 97.6 56.2 23.9 2 43.9 1.154.9 0.1 40.9 2.1 96.2 52.4 21.4 3 43.7 1.1 54.9 0.3 40.7 1.8 95.3 57.623.4 4 42.6 1.1 56 0.3 42.8 4.9 94.1 58.4 25.0 5 41.5 1.1 57 0.4 43.53.7 96.7 49.4 21.5 Comp. 1 41.1 1.1 57 0.8 43.7 2.3 89.4 30.5 13.3 Ex. 240.7 1.1 57 1.2 44.1 2.0 82.4 10.2 4.5

As is clear from the results shown in Table 1 and FIG. 1, thecompositions of the present invention (Examples 1 to 5) had highprotection and were also superior in dissolution.

On the other hand, the compositions of Comparative Examples 1 and 2showed low protection rate and low dissolution rate and were poor in invitro assumed efficacy rate compared to the control composition.

Experimental Example 2. Study of Kind of Natural Vegetable Oil Example 6

Soybean fully hydrogenated oil (manufactured by Yokozeki Oil & FatIndustries Co., Ltd., melting point: 67° C.), soybean lecithin(manufactured by ADM, Yelkin TS), L-lysine hydrochloride (manufacturedby Ajinomoto Co., Inc.) and olive oil (manufactured by JOM, extravirgin) in the proportions shown in the following Table 3-1 werecontinuously cast into a twin screw extruder (manufactured by CosmotecCo., Ltd.).

Thereafter, the mixture was heated (preheating temperature: 65° C., mainheating temperature: 85° C., set temperature for outlet: 70° C.), meltedand mixed in a cylinder to give a molten mixture in a molten slurrystate. The obtained molten mixture was discharged from the outlet of theextruder, cast into a multi-hole shooter (number of holes: 2060, holediameter: 2 mm), and the molten mixture was freely dropped from the holeof a multi-hole shooter into the water tank for cooling (watertemperature: 5 to 15° C.). The distance from the multi-hole shooter tothe water surface of the water tank for cooling was 10 cm. The moltenmixture that dropped from the multi-hole shooter became droplet duringdropping, immersed in water, cooled and solidified instantaneously. Theattached water was dehydrated by blowing at room temperature, and thesolid was subjected to a heat-drying treatment by a fluidized-bed dryer(manufactured by Ajinomoto Co., Inc.) set to 52° C. for 7 min to givegranules (feed additive composition for ruminants). In the following,the granulated product is referred to as the composition of Example 6.

Examples 7 to 12, and the Control

In the same manner as in Example 6 except that soybean oil (manufacturedby Wako Pure Chemical Industries, Ltd.), rape seed oil (manufactured byKaneda aburaten), avocado oil (manufactured by Kaneda aburaten), almondoil (manufactured by Kaneda aburaten), palm oil (manufactured by Kanedaaburaten) and safflower oil (manufactured by J-OIL MILLS, Inc.) in theproportions shown in the following Table 3-1 were used instead of oliveoil, the feed additive compositions for ruminants of Examples 7 to 11were respectively obtained (hereinafter to be respectively referred toas compositions of Examples 7 to 11).

In the same manner as in Example 6 except that the proportions shown inthe following Table 3-1 were used without using a natural vegetable oilsuch as olive oil and the like, the control feed additive compositionfor ruminants was obtained (hereinafter to be referred to as the controlcomposition).

The (i) content rate of unsaturated fatty acid having a carbon atomnumber of 18 to constituent fatty acid, (ii) content rate of oleic acidto constituent fatty acid, and (iii) unsaturation rate of fatty acidhaving a carbon atom number of 18 of olive oil, soybean oil, rape seedoil, avocado oil, almond oil, palm oil and safflower oil used forpreparing the compositions of Examples 6 to 12 are shown in thefollowing Table 2.

TABLE 2 Example 7 Example 8 Example 9 Example 12 Example 6 soybean rapeseed avocado Example 10 Example 11 safflower olive oil oil oil oilalmond oil palm oil oil (i) content rate 82.0 83.9 83.3 69.2 91.5 49.591.8 (wt %) of unsaturated fatty acid having carbon atom number of 18 toconstituent fatty acid (ii) content rate 72.8 25.7 58.0 55.8 65.6 39.678.0 (wt %) of oleic acid to constituent fatty acid (iii) unsaturationrate (%) of fatty 97.4 95.4 98.0 99.3 98.5 92.0 100 acid having carbonatom number of 18

The L-lysine content, water content, protection rate and dissolutionrate of the compositions of Examples 6 to 12 and the control weremeasured in the same manner as in Experimental Example 1. In addition,the in vitro assumed efficacy rate of the compositions of Examples 6 to12 and the control was calculated similarly to Experimental Example 1.

The L-lysine content, water content, protection rate, dissolution rateand in vitro assumed efficacy rate of the compositions of Examples 6 to12 and the control are shown in the following Table 3-2. In addition,the protection rate, dissolution rate and in vitro assumed efficacy rateof the compositions of Examples 6 to 12 and the control are shown inFIG. 2.

TABLE 3-1 blending proportion [wt %] (A) soybean fully (B) (C) (D) (D)(D) (D) (D) (D) (D) hydrogenated soybean L-lysine olive soybean rapeavocado almond palm safflower oil lecithin hydrochloride oil oil seedoil oil oil oil oil control 41.9 1.1 57 — — — — — — — Ex. 6 41.7 1.1 570.2 — — — — — — 7 41.7 1.1 57 — 0.2 — — — — — 8 41.7 1.1 57 — — 0.2 — —— — 9 41.7 1.1 57 — — — 0.2 — — — 10 41.7 1.1 57 — — — — 0.2 — — 11 41.71.1 57 — — — — — 0.2 — 12 41.7 1.1 57 — — — — — — 0.2

TABLE 3-2 L-lysine content water content [wt %] of protection in vitroassumed [wt %] of feed feed additive rate dissolution efficacy rateadditive composition composition [%] rate [%] [%] control 41.4 4.48 95.033.7 14.0 Ex. 6 43.3 3.83 90.8 53.7 23.3 7 42.4 3.63 89.9 53.6 22.7 844.0 3.85 91.1 49.7 21.9 9 43.6 3.69 90.3 48.7 21.3 10 44.2 3.86 89.752.7 23.3 11 43.0 3.67 91.8 47.6 20.5 12 43.1 4.61 97.3 59.2 25.5

As is clear from the results shown in Table 3-2 and FIG. 2, thecompositions of the present invention had high protection and were alsosuperior in dissolution when soybean oil, rape seed oil, avocado oil,almond oil, palm oil or safflower oil was used as Component D (naturalvegetable oil) (Examples 7 to 12), similar to the use of olive oil(Example 6).

Experimental Example 3. Study of Relationship Between Unsaturation Rateof Fatty Acid Having a Carbon Atom Number of 18 and In Vitro AssumedEfficacy Rate Reference Example

In the same manner as in Example 6 except that fully hydrogenatedsoybean oil (manufactured by Yokozeki Oil & Fat Industries Co., Ltd.,unsaturation rate of fatty acid having a carbon atom number of 18=0%)was used instead of olive oil, a feed additive composition for ruminantsof Reference Example (hereinafter to be referred to as the compositionof Reference Example) was obtained.

The protection rate and dissolution rate of the composition of ReferenceExample were measured in the same manner as in Experimental Example 1,and the in vitro assumed efficacy rate was calculated similarly toExperimental Example 1.

A graph plotting the in vitro assumed efficacy rate of the compositionsof Examples 6 to 12 and Reference Example relative to the unsaturationrate of fatty acid having a carbon atom number of 18 is shown in FIG. 3.

From the results shown in FIG. 3, it was confirmed that the unsaturationrate and in vitro assumed efficacy rate of the fatty acid having acarbon atom number of 18 in Component D (natural vegetable oil) arecorrelated, and as the unsaturation rate of fatty acid having a carbonatom number of 18 increases, in vitro assumed efficacy rate tends toincrease.

Experimental Example 4. Confirmation of Layer Substantially Free ofBiologically Active Substance

The composition of Example 1 obtained in Experimental Example 1 was cutand the section thereof was photographed with a scanning electronmicroscope (SEM). FIG. 4 shows an SEM photograph of the vicinity of thesurface of the composition of Example 1. From the photograph, it wasconfirmed that a layer smoothened by the heat treatment (layersubstantially free of a biologically active substance) was formed to adepth of 100 μm from the surface.

Experimental Example 5. Confirmation of Effect when Biologically ActiveSubstance is Histidine Hydrochloride Example 13 and Control

In the same manner as in Example 1 except that soybean fullyhydrogenated oil (manufactured by Yokozeki Oil & Fat Industries Co.,Ltd., melting point:67° C.), soybean lecithin (manufactured by ADM,Yelkin TS), L-histidine hydrochloride (manufactured by Ajinomoto Co.,Inc.) and olive oil (manufactured by JOM, extra virgin) were cast in theproportions shown in the following Table 4, feed additive compositionsfor ruminants of Example 13 and control were obtained (hereinafter to bereferred to as composition of Example 13 and control).

Measurement of L-Histidine Content of Feed Additive Composition

The L-histidine content of each composition was measured using liquidchromatography (manufactured by Hitachi).

The water content, protection rate and dissolution rate of thecompositions of Example 13 and control were measured in the same manneras in Experimental Example 1. In the measurement of protection rate anddissolution rate, the concentration of the biologically active substance(L-histidine) in the test solution was measured using liquidchromatography (manufactured by Hitachi). In addition, the in vitroassumed efficacy rate of the compositions of Example 13 and control wascalculated similarly to Experimental Example 1.

The L-histidine content, water content, protection rate, dissolutionrate and in vitro assumed efficacy rate of the compositions of Example13 and control are shown in the following Table 4. In addition, theprotection rate, dissolution rate and in vitro assumed efficacy rate ofthe compositions of Example 13 and control are shown in FIG. 5.

TABLE 4 blending proportion [wt %] L-histidine water (A) content contentsoybean [wt %] of [wt %] of in vitro fully (B) (C) (D) feed feedprotection dissolution assumed hydrogenated soybean L-histidine oliveadditive additive rate rate efficacy oil lecithin hydrochloride oilcomposition composition [%] [%] rate [%] control 38.7 1.3 60.0 0 42.50.34 98 9 3.8 Example 13 38.5 1.3 60.0 0.2 44.2 0.32 98 17 7.5

As is clear from the results shown in Table 4, the composition of thepresent invention (Example 13) had high protection and was superior indissolution.

On the other hand, the composition of the control showed low protectionrate and low dissolution rate.

Experimental Example 6. Confirmation of Effect when Biologically ActiveSubstance is Arginine Examples 14 and 15 and Control

In the same manner as in Example 1 except that soybean fullyhydrogenated oil (manufactured by Yokozeki Oil & Fat Industries Co.,Ltd., melting point:67° C.), soybean lecithin (manufactured by ADM,Yelkin TS), L-arginine (manufactured by Ajinomoto Co., Inc.) and oliveoil (manufactured by JOM, extra virgin) were cast in the proportionsshown in the following Table 5, feed additive compositions for ruminantsof Examples 14 and 15 and the control were obtained (hereinafter to bereferred to as composition of Examples 14 and 15 and control).

Measurement of L-Arginine Content of Feed Additive Composition

The L-arginine content of each composition was measured using liquidchromatography (manufactured by Hitachi).

The water content, protection rate and dissolution rate of thecompositions of Examples 14 and 15 and the control were measured in thesame manner as in Experimental Example 1. In addition, the in vitroassumed efficacy rates of the compositions of Examples 14 and 15 and thecontrol were calculated similarly to Experimental Example 1.

The L-arginine content, water content, protection rate, dissolution rateand in vitro assumed efficacy rate of the compositions of Examples 14and 15 and the control are shown in the following Table 5. In themeasurement of protection rate and dissolution rate, the concentrationof the biologically active substance (L-arginine) in the test solutionwas measured using liquid chromatography (manufactured by Hitachi). Inaddition, the protection rate, dissolution rate and in vitro assumedefficacy rate of the compositions of Examples 14 and 15 and the controlare shown in FIG. 6.

TABLE 5 L-arginine water blending proportion [wt %] content content (A)[wt %] of [wt %] of in vitro soybean fully (B) (C) (D) feed feedprotection dissolution assumed hydrogenated soybean L- olive additiveadditive rate rate efficacy oil lecithin arginine oil compositioncomposition [%] [%] rate [%] control 50.7 1.3 48 0 44.9 1.7 98 14.2 6.4Example 14 50.4 1.44 47.9 0.2 44.7 1.9 98 18.0 8.0 Example 15 50.3 1.3348 0.37 44.5 2.0 98 19.3 8.6

As is clear from the results shown in Table 5, the compositions of thepresent invention (Examples 14 and 15) had high protection and weresuperior in dissolution.

On the other hand, the composition of the control showed low in vitroassumed efficacy rate and low dissolution rate.

INDUSTRIAL APPLICABILITY

According to the present invention, a feed additive composition forruminants that is provided with high protection in the rumen and issuperior in dissolution in the gastrointestinal tract can be provided.

The feed additive composition for ruminants of the present inventioncontaining both lecithin and natural vegetable oil at particularconcentrations can further promote dissolution in the gastrointestinaltract while maintaining high protection in the rumen as compared to whenonly one of lecithin and natural vegetable oil is contained.

According to the feed additive composition for ruminants of the presentinvention, a large amount of a biologically active substance (e.g.,amino acid etc.) can be efficiently transported up to the smallintestine of lactating cow. Therefore, the lactating cow can absorb alarge amount of a biologically active substance (e.g., amino acid etc.)as a nutrient, as a result of which, for example, it is possible toincrease milk yield production and the like.

Where a numerical limit or range is stated herein, the endpoints areincluded. Also, all values and subranges within a numerical limit orrange are specifically included as if explicitly written out.

As used herein the words “a” and “an” and the like carry the meaning of“one or more.”

Obviously, numerous modifications and variations of the presentinvention are possible in light of the above teachings. It is thereforeto be understood that, within the scope of the appended claims, theinvention may be practiced otherwise than as specifically describedherein.

All patents and other references mentioned above are incorporated infull herein by this reference, the same as if set forth at length.

1. A feed additive composition, comprising: (A) at least one selectedfrom the group consisting of hydrogenated vegetable oil and hydrogenatedanimal oil each having a melting point of from higher than 50° C. tolower than 90° C.; (B) from 0.05 wt % to 6 wt %, based on the totalweight of the composition, of lecithin; (C) from 30 wt % to less than 65wt %, based on the total weight of the composition, of a biologicallyactive substance; (D) from 0.01 wt % to less than 0.8 wt %, based on thetotal weight of the composition, of a natural vegetable oil; and from0.1 wt % to less than 6 wt %, based on the total weight of thecomposition, of water.
 2. The composition according to claim 1, whereinthe amount of the natural vegetable oil (D) is from 0.1 wt % to 0.4 wt%, based on the total weight of the composition.
 3. The compositionaccording to claim 1, wherein the natural vegetable oil (D) is at leastone member selected from the group consisting of soybean oil, palm oil,rape seed oil, canola oil, olive oil, almond oil, avocado oil, andsafflower oil.
 4. The composition according to claim 2, wherein thenatural vegetable oil (D) is at least one member selected from the groupconsisting of soybean oil, palm oil, rape seed oil, canola oil, oliveoil, almond oil, avocado oil, and safflower oil.
 5. The compositionaccording to claim 1, wherein the natural vegetable oil (D) comprisesfrom 60 wt % to 95 wt % of an unsaturated fatty acid, relative to thetotal content of fatty acids in the natural vegetable oil (D), having 18carbon atoms.
 6. The composition according to claim 2, wherein thenatural vegetable oil (D) comprises from 60 wt % to 95 wt % of anunsaturated fatty acid, relative to the total content of fatty acids inthe natural vegetable oil (D), having 18 carbon atoms.
 7. Thecomposition according to claim 3, wherein the natural vegetable oil (D)comprises from 60 wt % to 95 wt % of an unsaturated fatty acid, relativeto the total content of fatty acids in the natural vegetable oil (D),having 18 carbon atoms.
 8. The composition according to claim 1, whereinthe natural vegetable oil (D) comprises from 55 wt % to 90 wt % of oleicacid, relative to the total content of fatty acids in the naturalvegetable oil (D).
 9. The composition according to claim 2, wherein thenatural vegetable oil (D) comprises from 55 wt % to 90 wt % of oleicacid, relative to the total content of fatty acids in the naturalvegetable oil (D).
 10. The composition according to claim 3, wherein thenatural vegetable oil (D) comprises from 55 wt % to 90 wt % of oleicacid, relative to the total content of fatty acids in the naturalvegetable oil (D).
 11. The composition according to claim 1, wherein thenatural vegetable oil (D) is olive oil.
 12. The composition according toclaim 2, wherein the natural vegetable oil (D) is olive oil.
 13. Thecomposition according to claim 3, wherein the natural vegetable oil (D)is olive oil.
 14. The composition according to claim 1, wherein thebiologically active substance (C) is at least one member selected fromthe group consisting of an amino acid, a vitamin, and a vitaminsubstance.
 15. The composition according to claim 2, wherein thebiologically active substance (C) is at least one member selected fromthe group consisting of an amino acid, a vitamin, and a vitaminsubstance.
 16. The composition according to claim 3, wherein thebiologically active substance (C) is at least one member selected fromthe group consisting of an amino acid, a vitamin, and a vitaminsubstance.
 17. The composition according to claim 1, wherein the meltingpoint of the component (A) is from 55° C. to 80° C.
 18. The compositionaccording to claim 1, wherein a content of the component (A) is from 30wt % to less than 60 wt %.
 19. The composition according to claim 1,wherein the feed additive composition has a form of particles having aparticle size of from 0.1 mm to 20 mm.
 20. The composition according toclaim 1, wherein the feed additive composition has a spherical,granular, pellet, rugby ball, pressed barley, of hen's egg shape.